# Isolation of F-specific Inovirus filamentous phages from environmental sewage samples

## TL;DR

This paper describes a simple method to isolate non-lytic filamentous phages from sewage using basic lab equipment.

## Contribution

A new protocol for isolating non-lytic F-specific inovirus phages from environmental samples is introduced.

## Key findings

- The protocol uses F-pilus expressing bacteria and sewage filtrate for phage enrichment.
- Hazy plaques were identified as potential filamentous phages and confirmed via PCR.
- The method is accessible for labs with basic microbiological tools.

## Abstract

Bacteriophage (phage) isolation from various environmental sources is an important step in identifying phages suitable for therapeutic and/or research purposes. Most published protocols outlining phage isolation techniques focus on the identification and/or characterization of strictly lytic phages. Here, a simple, adapted protocol specific for the isolation of non-lytic, filamentous phages is described. This method can be used in laboratories with access to basic microbiological equipment. Briefly, the adapted protocol steps involve:•Phage enrichment with F-pilus expressing bacterial host and sewage filtrate•Dilution and agar overlay plating of enriched culture•Selection of hazy, turbid plaques followed by screening via PCR

Phage enrichment with F-pilus expressing bacterial host and sewage filtrate

Dilution and agar overlay plating of enriched culture

Selection of hazy, turbid plaques followed by screening via PCR

Image, graphical abstract